Highly sensitive bioluminescence and fluorescence imaging system for large fields of view

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The major equipment will be used as an imaging system to read out transgenic fluorescent or bioluminescent reporter lines at the level of whole organs and organisms in molecular plant research. The applied for setup will enable imaging on a new scale, which will allow for the monitoring of systemic reactions to e.g. biotic or abiotic stress on the living sample in an organismic context. The processes studied are relevant for understanding the physiological responses of plants to abiotic and biotic stress conditions, as they increasingly occur with progressive climate change. The transgenic reporter lines used express e.g. genetically encoded biosensors (fluorescent proteins that change their properties specifically in response to a physiological parameter) with targeting to different subcellular compartments, or promoter or protein fusions with luciferase as reporter. This experimental approach allows monitoring of subcellular processes such as local changes in redox status or regulation of gene expression in a systemic context. With the major equipment applied for in this proposal, the bioluminescence or fluorescence of whole organisms such as plants can be recorded in large fields of view (approx. 20 cm). For this purpose, a high-performance CCD camera (16 bit) cooled to -90°C will be used, which has the necessary sensitivity and very low background noise (dark current). The camera setup is located in a light-proof box, which also contains the necessary illumination for fluorescence. To cover as many fluorescent proteins and genetically encoded biosensors as possible, suitable excitation and emission filters must be included. The adjustment of the instrument and acquisition parameters are controlled via a manufacturer-specific software from an external PC. The proposed device would be used for the first time in Germany in plant sciences.